摘要
根据苏云金芽胞杆菌(Bacillus thuringiensis)YBT-1520的全基因组测序结果,在内生质粒p BMB67和染色体上分别选取单拷贝片段,利用实时荧光定量PCR法测得质粒p BMB67的拷贝数为21。此外,在内生质粒p BMB67的3个不同位点设计引物,分别以不同的靶片段作为实时荧光定量PCR的检测对象,测得p BMB67质粒的拷贝数分别为20.1、21.0和21.3,表明单拷贝靶片段的选择对测定质粒的拷贝数没有影响,进一步证明了实时荧光定量PCR测定质粒拷贝数的可行性。
According to the whole genome sequence of Bacillus thuringiensis strain YBT-1520,a specific single-copy fragment was selected from the resident plasmid pBMB67 and chromosome separately. And the copy number （PCN） of pBMB67 was estimated as 21 by quantitative real-time PCR. In addition,three pairs of primers were designed at three different fragments of pBMB67 as the targets of quantitative real-time PCR detection,and the result showed that the PCN of pBMB67 was 21.3,20.1, and 21.0,respectively. This finding indicated that the difference of target fragments on plasmid for quantitative real-time PCR detection would not affect the value of PCN,and demonstrated that the quantitative real-time PCR method is reliable to determine the PCN.
出处
《湖北农业科学》
2015年第8期2001-2006,共6页
Hubei Agricultural Sciences
基金
湖北省自然科学基金项目(2014CFB566)
